Abstract
The epidermis is largely composed of keratinocytes (KCs), and the proliferation and differentiation of KCs from the stratum basale to the stratum corneum is the cellular hierarchy present in the epidermis. In this study, we explore the differentiation abilities of human hematopoietic stem cells (HSCs) into KCs. Cultured HSCs positive for CD34, CD45, and CD133 with prominent telomerase activity were induced with keratinocyte differentiation medium (KDM), which is composed of bovine pituitary extract (BPE), epidermal growth factor (EGF), insulin, hydrocortisone, epinephrine, transferrin, calcium chloride (CaCl2), bone morphogenetic protein 4 (BMP4), and retinoic acid (RA). Differentiation was monitored through the expression of cytokeratin markers K5 (keratin 5), K14 (keratin 14), K10 (keratin 10), K1 (keratin 1), transglutaminase 1 (TGM1), involucrin (IVL), and filaggrin (FLG) on day 0 (D0), day 6 (D6), day 11 (D11), day 18 (D18), day 24 (D24), and day 30 (D30) using immunocytochemistry, fluorescence microscopy, flow cytometry, qPCR, and Western blotting. The results revealed the expression of K5 and K14 genes in D6 cells (early keratinocytes), K10 and K1 genes in D11–D18 cells (mature keratinocytes) with active telomerase enzyme, and FLG, IVL, and TGM1 in D18–D24 cells (terminal keratinocytes), and by D30, the KCs were completely enucleated similar to cornified matrix. This method of differentiation of HSCs to KCs explains the cellular order exists in the normal epidermis and opens the possibility of exploring the use of human HSCs in the epidermal differentiation.
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Acknowledgements
We sincerely acknowledge the Sri Venkateswara Institute of Medical Sciences (SVIMS University), Tirupati, India, for providing the facilities to carry out this work and this paper forms a part of a Ph.D. thesis work to be submitted/submitted to SVIMS University, Tirupati, Andhra Pradesh, India.
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This work was supported by the Sri Balaji Arogya Vara Prasadini Scheme (AS/01/MRC/SVIMS/2021-dated 08–02-2022) from the Sri Venkateswara Institute of Medical Sciences, Tirupati, Andhra Pradesh. The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.
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PVGK Sarma: visualization, investigation, reviewing, and editing. Kodavala Sireesha: conceptualization, methodology, and writing—original draft preparation. Echambadi Loganathan Samundeshwari: methodology and resources. Kattaru Surekha: methodology and resources. Chodimella Chandrasekhar: reviewing and editing.
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Sireesha, K., Samundeshwari, E.L., Surekha, K. et al. In vitro generation of epidermal keratinocytes from human CD34-positive hematopoietic stem cells. In Vitro Cell.Dev.Biol.-Animal 60, 236–248 (2024). https://doi.org/10.1007/s11626-024-00862-x
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DOI: https://doi.org/10.1007/s11626-024-00862-x